Transferrin-derived synthetic peptide induces highly conserved pro-inflammatory responses of macrophages.
We examined the induction of macrophage pro-inflammatory responses by transferrin-derived synthetic peptide originally identified following digestion of transferrin from different species (murine, bovine, human N-lobe and goldfish) using elastase. The mass spectrometry analysis of elastase-digested murine transferrin identified a 31 amino acid peptide located in the N2 sub-domain of the transferrin N-lobe, that we named TMAP. TMAP was synthetically produced and shown to induce a number of pro-inflammatory genes by quantitative PCR. TMAP induced chemotaxis, a potent nitric oxide response, and TNF-alpha secretion in different macrophage populations; P338D1 macrophage-like cells, mouse peritoneal macrophages, mouse bone marrow-derived macrophages (BMDM) and goldfish macrophages. The treatment of BMDM cultures with TMAP stimulated the production of nine cytokines and chemokines (IL-6, MCP-5, MIP-1 alpha, MIP-1 gamma, MIP-2, GCSF, KC, VEGF, and RANTES) that was measured using cytokine antibody array and confirmed by Western blot. Our results indicate that transferrin-derived peptide, TMAP, is an immunomodulating molecule capable of inducing pro-inflammatory responses in lower and higher vertebrates.
Haddad
George
G
Department of Biological Sciences, University of Alberta, CW-405 Biological Sciences Building, Edmonton, AB, T6G 2E9 Canada.
Belosevic
Miodrag
M
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002633:Chemotaxis / Q000276:immunology; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006054:Goldfish; D006801:Humans; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000276:immunology / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism*; D014168:Transferrin / Q000276:immunology / Q000378:metabolism*
D016207:Cytokines; D010455:Peptides; D014168:Transferrin; D009569:Nitric Oxide